Direct venous inoculation of Plasmodium falciparum sporozoites for controlled human malaria infection: a dose-finding trial in two centres.

BACKGROUND: Controlled human malaria infection (CHMI) accelerates development of anti-malarial interventions. So far, CHMI is done by exposure of volunteers to bites of five mosquitoes carrying Plasmodium falciparum sporozoites (PfSPZ), a technique available in only a few centres worldwide. Mosquito-mediated CHMI is logistically complex, exact PfSPZ dosage is impossible and live mosquito-based interventions are not suitable for further clinical development. METHODS: An open-labelled, randomized, dose-finding study in 18-45 year old, healthy, malaria-naïve volunteers was performed to assess if intravenous (IV) injection of 50 to 3,200 aseptic, purified, cryopreserved PfSPZ is safe and achieves infection kinetics comparable to published data of mosquito-mediated CHMI. An independent study site verified the fully infectious dose using direct venous inoculation of PfSPZ. Parasite kinetics were assessed by thick blood smear microscopy and quantitative real time PCR. RESULTS: IV inoculation with 50, 200, 800, or 3,200 PfSPZ led to parasitaemia in 1/3, 1/3, 7/9, and 9/9 volunteers, respectively. The geometric mean pre-patent period (GMPPP) was 11.2 days (range 10.5-12.5) in the 3,200 PfSPZ IV group. Subsequently, six volunteers received 3,200 PfSPZ by direct venous inoculation at an independent investigational site. All six developed parasitaemia (GMPPP: 11.4 days, range: 10.4-12.3). Inoculation of PfSPZ was safe. Infection rate and pre-patent period depended on dose, and injection of 3,200 PfSPZ led to a GMPPP similar to CHMI with five PfSPZ-infected mosquitoes. The infectious dose of PfSPZ predicted dosage of radiation-attenuated PfSPZ required for successful vaccination. CONCLUSIONS: IV inoculation of PfSPZ is safe, well tolerated and highly reproducible. It shall further accelerate development of anti-malarial interventions through standardization and facilitation of CHMI. Beyond this, rational dose selection for whole PfSPZ-based immunization and complex study designs are now possible. TRIAL REGISTRATION: ClinicalTrials.gov NCT01624961 and NCT01771848 .

Delayed hemolysis after treatment with parenteral artesunate in African children with severe malaria--a double-center prospective study.

BACKGROUND: Parenteral artesunate is recommended as first-line therapy for severe malaria. While its efficacy is firmly established, data on safety are still incomplete. Delayed hemolysis has been described in hyperparasitemic nonimmune travelers, but it is unknown if African children are equally at risk. METHODS: Children aged 6 to 120 months with severe malaria were followed up after treatment with parenteral artesunate in Lambaréné, Gabon, and Kumasi, Ghana. The primary outcome was incidence of delayed hemolysis on day 14. RESULTS: In total, 72 children contributed complete data sets necessary for primary outcome assessment. Delayed hemolysis was detected in 5 children (7%), with 1 child reaching a nadir in hemoglobin of 2.8 g/dL. Patients with delayed hemolysis had higher parasite counts on admission (geometric mean parasite densities (GMPD) 306 968/µL vs 92 642/µL, P = .028) and were younger (median age: 24 months vs 43 months, P = .046) than the rest of the cohort. No correlation with sickle cell trait or glucose-6-phosphate-dehydrogenase deficiency was observed. CONCLUSIONS: Delayed hemolysis is a frequent and relevant complication in hyperparasitemic African children treated with parenteral artesunate for severe malaria. Physicians should be aware of this complication and consider prolonged follow-up. CLINICAL TRIALS REGISTRATION: Pan-African Clinical Trials Registry: PACTR201102000277177 (www.pactr.org).

Limit of blank and limit of detection of Plasmodium falciparum thick blood smear microscopy in a routine setting in Central Africa.

BACKGROUND: Proper malaria diagnosis depends on the detection of asexual forms of Plasmodium spp. in the blood. Thick blood smear microscopy is the accepted gold standard of malaria diagnosis and is widely implemented. Surprisingly, diagnostic performance of this method is not well investigated and many clinicians in African routine settings base treatment decisions independent of microscopy results. This leads to overtreatment and poor management of other febrile diseases. Implementation of quality control programmes is recommended, but requires sustained funding, external logistic support and constant training and supervision of the staff. This study describes an easily applicable method to assess the performance of thick blood smear microscopy by determining the limit of blank and limit of detection. These two values are representative of the diagnostic quality and allow the correct discrimination between positive and negative samples. METHODS: Standard-conform methodology was applied and adapted to determine the limit of blank and the limit of detection of two thick blood smear microscopy methods (WHO and Lambaréné method) in a research centre in Lambaréné, Gabon. Duplicates of negative and low parasitaemia thick blood smears were read by several microscopists. The mean and standard deviation of the results were used to calculate the limit of blank and subsequently the limit of detection. RESULTS: The limit of blank was 0 parasites/µL for both methods. The limit of detection was 62 and 88 parasites/µL for the Lambaréné and WHO method, respectively. CONCLUSION: With a simple, back-of-the-envelope calculation, the performance of two malaria microscopy methods can be measured. These results are specific for each diagnostic unit and cannot be generalized but implementation of a system to control microscopy performance can improve confidence in parasitological results and thereby strengthen malaria control.